[0001] The present invention relates to pharmaceuticals. Specifically, the present invention relates to geranyl geranyl transferase inhibitoras therapeutic target fordiabetic liver injury. The present invention further relates to a specific dosage of geranyl geranyl transferase inhibitor as hepatoprotective for diabetic liver injury.

BACKGROUND OF THE INVENTION
[0002] Background description includes information that may be useful in understanding the present invention. It is not an admission that any of the information provided herein is prior art or relevant to the presently claimed invention, or that any publication specifically or implicitly referenced is prior art.
[0003] Diabetes is associated with a spectrum of liver diseases including non-alcoholic fatty liver disease (NAFLD), steatohepatitis, and liver cirrhosis with their increased complications and mortality (Musso et al, 2011). The epidemiological studies reported that the occurrence of NAFLD in type 2 diabetes mellitus (T2DM) patients ranges from 50% to 75% and in India the prevalence range is reported to be 12.5-87.5% (Kalara et al., 2013; Younossi et al., 2019). NAFLD has been recognized as a common complication in patients with T2DM.High fructose and fat intake elevate serum concentration of free fatty acids which result in accumulation of triglycerides, long-chain acyl-coenzyme A and DAG in hepatocytes and induce a diabetic like condition (Zhao et al., 2016).High fructose high fat (HFHF) diet generates free radical species, disturb triglycerides clearance from blood circulation and promote the development of various metabolic syndromes (Nakagawa et al., 2006).Consumption of large amount of western diet i.e., diet rich in fat and fructose is increasing chronically in today’s world which leads to development of diabetic associate liver injury. High fructose high fat diet generates free radical species, disturb triglycerides clearance from blood circulation and promote the development of various metabolic syndromes (Nakagawa et al., 2006).
[0004] Existing drug therapies help in the treatment of the disease by increasing fatty acid oxidation, suppressing the expression of lipogenic enzymes, improving glucose metabolism, increasing plasma adiponectin (Ibrahim et al., 2013).
[0005] Development of preventive measures to get rid from this chronic disease by studying dual model of diabetes and liver injury is an important public health concern. Although therapeutic solutions for NAFLD are already on the market, new therapeutic solutions are urgently required to address the growing number of NAFLD cases experienced by today’s society. There is, therefore, a need to develop novel compounds that are capable of acting as therapeutic solutions for non-alcoholic fatty liver disease and can overcome deficiencies associated with the known arts.

OBJECTS OF THE INVENTION
[0006] An object of the present invention is to provide novel therapeutic target for diabetic liver injury that can overcome deficiencies associated with the known arts.
[0007] An object of the present invention is to provide geranyl geranyl transferase inhibitor as therapeutic targets for diabetic liver injury.
[0008] Yet another object of the present invention is to provide specific dosage of geranyl geranyl transferase inhibitor as therapeutic targets for diabetic liver injury.
[0009] Another object of the present invention is to provide geranyl geranyl transferase inhibitor that is hepatoprotective, safe to use and has no side effects.
[0010] Yet another object of the present invention is to provide geranyl geranyl transferase inhibitor that reduces blood sugar and body weight gain associated with diabetic liver injury.
[0011] The other objects and preferred embodiments and advantages of the present invention will become more apparent from the following description of the present invention when read in conjunction with the accompanying examples and figures, which are not intended to limit scope of the present invention in any manner.

SUMMARY OF THE INVENTION
[0012] This summary is provided to introduce a selection of concepts in a simplified form that are further described below in Detailed Description section. This summary is not intended to identify key features or essential features of the claimed subject matter, nor is it intended to be used as an aid in determining the scope of the claimed subject matter.
[0013] The present invention relates to pharmaceuticals. Specifically, the present invention relates to geranyl geranyl transferase inhibitor as therapeutic target for diabetic liver injury.
[0014] In one aspect, the present invention relates to geranyl geranyl transferase inhibitor GGTI-2133 having hepatoprotective effect for diabetic liver injury.
[0015] In one aspect, the present invention relates to daily dose of GGTI-2133 which exerts a hepatoprotective effect against diabetic liver injury.
[0016] In one aspect, the present invention relates to GGTI-2133, wherein the daily dose of GGTI-2133 for hepatoprotective effect against diabetic liver injury is in the range of 0.001mg/kg to 5 mg/kg.
[0017] Various objects, features, aspects and advantages of the inventive subject matter will become more apparent from the following detailed description of preferred embodiments.

BRIEF DESCRIPTION OF DRAWINGS THE INVENTION
[0018] The following drawings form part of the present specification and are included to further illustrate aspects of the present disclosure. The disclosure may be better understood by reference to the drawings in combination with the detailed description of the specific embodiments presented herein.
Figure 1: Effects of selected pharmacological intervention on body weight changes in various experimental rodent groups.
*P < 0.05 versus control; @P < 0.05 versus Disease control HFHF.
Figure 2: Effects of selected pharmacological intervention on blood glucose levels in various experimental rodent groups.
*P < 0.05 versus control; @P < 0.05 versus Disease control HFHF
Figure 3: Effects of selected pharmacological intervention on liver function parameters levels in various experimental rodent groups.
*P < 0.05 versus control; @P < 0.05 versus Disease control HFHF.
Figure 4: Effects of selected pharmacological intervention on lipid profile parameter (HDL) levels in various experimental rodent groups.
*P < 0.05 versus control; @P < 0.05 versus Disease control HFHF; @P < 0.05 versus GGTI (0.1mg/Kg).
Figure 5: Effects of selected pharmacological intervention on lipid profile parameters levels in various experimental rodent groups.*P < 0.05 versus control; @P < 0.05 versus Disease control HFHF; #P < 0.05 versus Standard MET (120mg/kg); $P < 0.05 versus GGTI (0.1mg/Kg).
Figure 6: Effects of selected pharmacological intervention on TBARS levels in various experimental rodent groups.
*P < 0.05 versus control; @P < 0.05 versus Disease control HFHF.
Figure 7: Effects of selected pharmacological intervention reduced glutathione levels in various experimental rodent groups.
*P < 0.05 versus control; @P < 0.05 versus Disease control HFHF.
Figure 8: Effect of selected pharmacological intervention on serum free fatty acid levels level levels in various experimental rodent groups.
*P < 0.05 versus control; @P < 0.05 versus Disease control HFHF; #P < 0.05 versus Standard MET (120mg/kg); $P < 0.05 versus GGTI (0.1mg/Kg).

DETAILED DESCRIPTION
[0019] The following is a detailed description of embodiments of the disclosure. The embodiments are in such detail as to clearly communicate the disclosure. However, the amount of detail offered is not intended to limit the anticipated variations of embodiments; on the contrary, the intention is to cover all modifications, equivalents, and alternatives falling within the spirit and scope of the present disclosure as defined by the appended claims.
[0020] All publications herein are incorporated by reference to the same extent as if each individual publication or patent application were specifically and individually indicated to be incorporated by reference. Where a definition or use of a term in an incorporated reference is inconsistent or contrary to the definition of that term provided herein, the definition of that term provided herein applies and the definition of that term in the reference does not apply.
[0021] Reference throughout this specification to “one embodiment” or “an embodiment” means that a particular feature, structure or characteristic described in connection with the embodiment is included in at least one embodiment. Thus, the appearances of the phrases “in one embodiment” or “in an embodiment” in various places throughout this specification are not necessarily all referring to the same embodiment. Furthermore, the particular features, structures, or characteristics may be combined in any suitable manner in one or more embodiments.
[0022] In some embodiments, the numbers expressing quantities of ingredients, properties such as concentration, reaction conditions, and so forth, used to describe and claim certain embodiments of the invention are to be understood as being modified in some instances by the term “about.”Accordingly, in some embodiments, the numerical parameters set forth in the written description and attached claims are approximations that can vary depending upon the desired properties sought to be obtained by a particular embodiment. In some embodiments, the numerical parameters should be construed in light of the number of reported significant digits and by applying ordinary rounding techniques. Notwithstanding that the numerical ranges and parameters setting forth the broad scope of some embodiments of the invention are approximations, the numerical values set forth in the specific examples are reported as precisely as practicable. The numerical values presented in some embodiments of the invention may contain certain errors necessarily resulting from the standard deviation found in their respective testing measurements.
[0023] As used in the description herein and throughout the claims that follow, the meaning of “a,” “an,” and “the” includes plural reference unless the context clearly dictates otherwise. Also, as used in the description herein, the meaning of “in” includes “in” and “on” unless the context clearly dictates otherwise.
[0024] Unless the context requires otherwise, throughout the specification which follow, the word “comprise” and variations thereof, such as, “comprises” and “comprising” are to be construed in an open, inclusive sense that is as “including, but not limited to.”
[0025] The recitation of ranges of values herein is merely intended to serve as a shorthand method of referring individually to each separate value falling within the range. Unless otherwise indicated herein, each individual value is incorporated into the specification as if it were individually recited herein. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g. “such as”) provided with respect to certain embodiments herein is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention otherwise claimed. No language in the specification should be construed as indicating any non-claimed element essential to the practice of the invention.
[0026] Groupings of alternative elements or embodiments of the invention disclosed herein are not to be construed as limitations. Each group member can be referred to and claimed individually or in any combination with other members of the group or other elements found herein. One or more members of a group can be included in, or deleted from, a group for reasons of convenience and/or patentability. When any such inclusion or deletion occurs, the specification is herein deemed to contain the group as modified thus fulfilling the written description of all Markush groups used in the appended claims.
[0027] The description that follows, and the embodiments described therein, is provided by way of illustration of an example, or examples, of particular embodiments of the principles and aspects of the present disclosure. These examples are provided for the purposes of explanation, and not of limitation, of those principles and of the disclosure.
[0028] It should also be appreciated that the present disclosure can be implemented in numerous ways, including as a system, a method or a device. In this specification, these implementations, or any other form that the invention may take, may be referred to as processes. In general, the order of the steps of the disclosed processes may be altered within the scope of the invention.
[0029] The headings and abstract of the invention provided herein are for convenience only and do not interpret the scope or meaning of the embodiments.
[0030] The following discussion provides many example embodiments of the inventive subject matter. Although each embodiment represents a single combination of inventive elements, the inventive subject matter is considered to include all possible combinations of the disclosed elements. Thus if one embodiment comprises elements A, B, and C, and a second embodiment comprises elements B and D, then the inventive subject matter is also considered to include other remaining combinations of A, B, C, or D, even if not explicitly disclosed.
[0031] Various terms as used herein are shown below. To the extent a term used in a claim is not defined below, it should be given the broadest definition persons in the pertinent art have given that term as reflected in printed publications and issued patents at the time of filing.
[0032] The present invention relates to pharmaceuticals. Specifically, the present invention relates to geranyl geranyl transferase inhibitor as therapeutic target for diabetic liver injury.
[0033] In one embodiment, the present invention relates to geranyl geranyl transferase inhibitor GGTI-2133 having hepatoprotective effect for diabetic liver injury.
[0034] Geranyl geranyl transferase (GGTase) is noted to mediate a number of signal transduction cascades which are known to be involved in the causation of insulin resistance and hepatocellular carcinoma. GGTI-2133 is selective modulator of geranyl geranyl transferase.
[0035] According to embodiments of the present invention, GGTI-2133 acts as GGTase inhibitor for the management and prevention of HFHF induced diabetic liver injury, particularly in rodents.
[0036] The chemical name of GGTI-2133 is N-[[4-(Imidazol-4-yl)methylamino]-2-(1-naphthyl)benzoyl]leucine trifluoroacetate salt and its chemical structure is depicted below:

[0037] In one embodiment, the present invention relates to daily dose of GGTI-2133 which exerts a hepatoprotective effect against diabetic liver injury.
[0038] In one embodiment, the present invention relates to GGTI-2133, wherein the daily dose for hepatoprotective effect against diabetic liver injury is in the range of 0.001mg/kg to 5 mg/kg.
[0039] In a preferred embodiment, the present invention relates to GGTI-2133, wherein the daily dose for hepatoprotective effect against diabetic liver injury is in the range of 0.01mg/kg to 5 mg/kg.
[0040] In a most preferred embodiment, the present invention relates to GGTI-2133, wherein the daily dose for hepatoprotective effect against diabetic liver injury is in the range of 0.1mg/kg to 0.3 mg/kg.
[0041] According to embodiments of the present invention,GGTI-2133, at a daily dose of 0.1mg/kg and/or 0.3mg/kg exerts a hepatoprotective effect against diabetic liver injury by inhibiting protein prenylation in cholesterol synthesis pathway (mevalonate pathway) and reducing oxidative stress, lipid profile, liver profile and FFA levels.
[0042] In another embodiment, the present invention relates to GGTI-2133 which at a daily dosage of 0.1mg/kg and/or 0.3mg/kg exerts a hepatoprotective effect against diabetic liver injury, particularly in rats. The results obtained for GGTI-2133 in rats can be extrapolated to human dosages by carrying out further experiments and discloses its potential beneficial role for patients with diabetic liver injury.
[0043] While the foregoing describes various embodiments of the disclosure, other and further embodiments of the disclosure may be devised without departing from the basic scope thereof. The scope of the invention is determined by the claims that follow. The invention is not limited to the described embodiments, versions or examples, which are included to enable a person having ordinary skill in the art to make and use the invention when combined with information and knowledge available to the person having ordinary skill in the art.
[0044] The present invention is further explained in the form of following examples. However, it is to be understood that the following examples are merely illustrative and are not to be taken as limitations upon the scope of the invention.
[0045] GGTI-2133 (Sigma Aldrich) was purchased from local vendor VS corporation Chandigarh.
[0046] Example1:Animal Models
[0047] Wistar rats of one month old weighing 100-120gm were employed in the present study.Diabetic liver injury was induced by consumption of high fat high fructose diet (HFHF) for approximately seven months (as described in Lozano et al., 2016). Blood glucose level was measured using glucometer and body weights were measured during the study on specific time intervals. Rats were randomly assigned into five groups each comprising of 6 rats as shown below:
[0048] Group I (Control group), animals were exposed to normal diet (seven months) during the experiment and blood glucose level and body weight are measured during the experiment.
[0049] Group II (Disease control group), animals were exposed to high fat high fructose diet for seven months. During the period their body weight, blood glucose level were increased abruptly.
[0050] Group III (Standard metformin treated group) after the induction of diabetic liver injury (25th week), when blood glucose level of rodents was increased, they were treated with standard metformin (120mg/kg) IP.
[0051] Group IV (GGTI-2133, 0.1 mg/kg treated group) after the induction of the DLI (25th week) when the blood glucose level of rodents was above 200mg/dl they were treated with 0.1mg/kg IP injection of Gernayl Gernayl Transferase-2133 (GGTI-2133).
[0052] Group V (GGTI-2133, 0.3 mg/kg treated group) after the induction of the DLI (25th week) when the blood glucose level of rodents was increased, they were treated with 0.3mg/kg IP injection of Gernayl Gernayl Transferase-2133 (GGTI-2133).
[0053] Research diet Formula: The diet described in Table 1 was used in the experiments.
Table 1
Product % Gms Kcal
Protein 19.3 20
Carbohydrate 67.5 70
Fat 4.3 10
Total 91.1 100

Ingredients
Casein, 80 Mesh 200 800
L-Cystein 3 12
Fructose 710 2840
Cellulose, BW200 50 0
Soyabean oil 25 225
Lard 20 180
Mineral Mix S10040A 6.00895 0
Mineral Mix S10026 0 0
DiCalcium Phosphate 13 0

[0054] Assessment of TBARS and reduced glutathione was performed by standard procedures as described in Kumar and Kalonia, 2007.
[0055] Assessment of liver profile (AST, ALT and ALP levels) were carried out using commercial biochemical enzymatic diagnostic kit as described in Arthurs et al., 1973; Lieberman and Phillips, 1990.
[0056] Assessment of Free fatty Acids was done by using standard commercial diagnostic kits.
[0057] Assessment of lipid profile: The quantifications of lipid profile was done by using standard commercial diagnostic kits as per the procedures of Russell Warnick et al., 1982, and Henkel et al., 1982.
[0058] Statistical Analysis: Statistical analysis was carried out by using appropriate software (Graph pad prism or Sigma stat).
[0059] Example 2: Biochemical Estimation and Results
[0060] Effect of GGTI-2133 treatments on blood glucose level and body weight in the rodent model of diabetic liver injury
[0061] Body weight: Administration of HFHF diet for 7 months to rodents shown a significant (P<0.001) increase in body weight as compared to vehicle treated experimental group, measured on 28th week during experimentation. The GGTI-2133 treated groups (0.1 and 0.3 mg/kg IP) shows significant decrease in body weight as compared to HFHF diet treated group on 28th week. Standard met for min and 0.1mg/kg GGTI-2133 treated group also shows significant decrease in body weight when compared to disease control group. These results are described in Figure 1.
[0062] Blood glucose: It was observed that disease control group (HFHF) of rodent models of diabetic liver injury show a significantly (P<0.001) higher blood glucose level when compared to vehicle treated experimental group measured on 28th week during experimentation. It was also observed that 0.3mg/kg GGTI-2133, 0.1mg/kg GGTI-2133 and standard met for min treated groups shown significant decreased blood glucose level as compared to disease control group of rodent models of Diabetic liver injury. These results are described in Figure 2.

[0063] Effect of GGTI-2133 on liver enzymes AST and ALT and ALP in rodent models of HFHF diet induced diabetic liver injury
[0064] It was found that oral administration of high fat high fructose diet causes significant (P < 0.001) increase in AST and ALT and ALP levels in disease control group as compared to vehicle treated group of rodent models of diabetic liver injury. 0.3 mg/kg GGTI-2133 shows a significant decrease in AST and ALT level when compared to disease control group (HFHF). It was also observed that GGTI-2133 at 0.3mg/kg shows significant (P < 0.05) dose dependent decrease in AST and ALT level as compared to 0.1mg/kg GGTI-2133 treated group. Standard metformin and 0.1mg/kg GGTI-2133 also showed significant decrease in AST and ALT level as compared to disease control group. These results are described in Figure3.
[0065] Effect of GGTI-2133 treatments on high density lipoprotein (HDL) levels in rodent’s model of diabetic liver injury
[0066] It was observed that there was significant (P < 0.001) decrease in HDL level in disease control group of high fat high fructose induced diabetic liver injury as compared to vehicle treated experimental group. Also, there was significant increase in HDL level in 0.3mg/kg GGTI-2133 and standard metformin treated group as compared to disease control group of HFHF diet induced diabetic liver injury. It was also observed that GGTI 0.3mg/kg shows significant (P < 0.05) dose dependent increase in HDL level as compared to 0.1mg/kg GGTI treated group. These results are described in Figure4.
[0067] Effect of GGTI-2133 treatments on LDL, triglycerides and total cholesterol levels in rodent’s model of diabetic liver injury
[0068] It was found that there was significant (P < 0.001) increase in LDL, triglycerides and cholesterol levels in disease control group as compared to vehicle treated group of Diabetic liver injury. It was also observed that there was significant decrease in LDL, triglycerides and cholesterol levels in 0.3mg/kg GGTI treated group as compared to disease control (HFHF) group of diabetic liver injury. Decreased level of LDL, triglycerides and cholesterol was observed in 0.3mg/kg GGTI-2133 treated group in comparison with standard metformin group. It was also observed that GGTI 0.3mg/kg shown significant (P < 0.05) dose dependent decrease in LDL level as compared to 0.1mg/kg GGTI-2133 treated group. These results are described in Figure5.
[0069] Effect of GGTI-2133 treatment on various biochemical estimation of HFHF induced Diabetic Liver Injury
[0070] Thiobarbituric acid reactive substances (TBARS): The level of lipid peroxidation in liver, kidney was studied by measuring the Thiobarbituric acid reactive substances (TBARS) in the liver, kidney homogenate method. There was significant (P < 0.001) increase in TBARS levels in disease control group when compared to vehicle treated group. Treatment with 0.3mg/kg GGTI-2133 reduced the level of TBARS when compared to disease control group. It was also observed that 0.1mg/kg GGTI-2133 treated group and standard metformin treated group shown significant decrease in TBARS level when compared to disease control group. These results are described in Figure6.
[0071] Effect of GGTI-2133 treatments on Glutathione
[0072] It was observed that the oral administration of high fat high fructose diet for seven months significantly (P < 0.001) decreased the level of reduced glutathione in disease control group of diabetic liver injury as compared to vehicle treated group of rodents. There was significant increase in level of reduced glutathione in 0.3 mg/kg GGTI-2133 treated group as compared to disease control group of HFHF diet induced diabetic liver injury group. Standard metformin and 0.1mg/kg GGTI-2133 treated groups also shows increased level of reduced glutathione as compared to disease control group. These results are described in Figure7.
[0073] Effect of GGTI-2133 treatments on Free fatty acid
[0074] The level of free fatty acid was significantly (P < 0.001) higher in disease control model of High fat high fructose diet induce diabetic liver injury as compared to vehicle treated experimental group. There was significant reduction of FFA level in 0.3mg/kg GGTI-2133 treated group as compared to disease control group. FFA level in 0.3mg/kg GGTI-2133 treated group was also reduced as compared to standard metformin treated group. It was also observed that GGTI 0.3mg/kg show significant (P < 0.05) dose dependent decrease in free fatty acid level when compared to 0.1mg/kg GGTI treated group. These results are described in Figure 8.
[0075] The foregoing examples are merely illustrative and are not to be taken as limitations upon the scope of the invention. Various changes and modifications to the disclosed embodiments will be apparent to those skilled in the art. Such changes and modifications may be made without departing from the scope of the invention.

Claims:1. Dose of geranyl geranyl transferase inhibitor as hepatoprotective against diabetic liver injury, wherein the daily dose is in the range of 0.001mg/kg to 5 mg/kg and geranyl geranyl transferase inhibitor is GGTI-2133.
2. The dose as claimed in claim 1, wherein the dose is in the range of 0.01mg/kg to 5 mg/kg.
3. The dose as claimed in claim 1, wherein the dose is in the range of 0.1mg/kg to 0.3 mg/kg.